Clinical utility of whole-genome DNA methylation profiling as a primary molecular diagnostic assay for central nervous system tumors-A prospective study and guidelines for clinical testing.

Background: Central nervous system (CNS) cancer is the 10th leading cause of cancer-associated deaths for adults, but the leading cause in pediatric patients and young adults. The variety and complexity of histologic subtypes can lead to diagnostic errors. DNA methylation is an epigenetic modification that provides a tumor type-specific signature that can be used for diagnosis. Methods: We performed a prospective study using DNA methylation analysis as a primary diagnostic method for 1921 brain tumors. All tumors received a pathology diagnosis and profiling by whole genome DNA methylation, followed by next-generation DNA and RNA sequencing. Results were stratified by concordance between DNA methylation and histopathology, establishing diagnostic utility. Results: Of the 1602 cases with a World Health Organization histologic diagnosis, DNA methylation identified a diagnostic mismatch in 225 cases (14%), 78 cases (5%) did not classify with any class, and in an additional 110 (7%) cases DNA methylation confirmed the diagnosis and provided prognostic information. Of 319 cases carrying 195 different descriptive histologic diagnoses, DNA methylation provided a definitive diagnosis in 273 (86%) cases, separated them into 55 methylation classes, and changed the grading in 58 (18%) cases. Conclusions: DNA methylation analysis is a robust method to diagnose primary CNS tumors, improving diagnostic accuracy, decreasing diagnostic errors and inconclusive diagnoses, and providing prognostic subclassification. This study provides a framework for inclusion of DNA methylation profiling as a primary molecular diagnostic test into professional guidelines for CNS tumors. The benefits include increased diagnostic accuracy, improved patient management, and refinements in clinical trial design.

Clinical Validation of Stimulated Raman Histology for Rapid Intraoperative Diagnosis of Central Nervous System Tumors.

Stimulated Raman histology (SRH) is an ex vivo optical imaging method that enables microscopic examination of fresh tissue intraoperatively. The conventional intraoperative method uses frozen section analysis, which is labor and time intensive, introduces artifacts that limit diagnostic accuracy, and consumes tissue. SRH imaging allows rapid microscopic imaging of fresh tissue, avoids tissue loss, and enables remote telepathology review. This improves access to expert neuropathology consultation in both low- and high-resource practices. We clinically validated SRH by performing a blinded, retrospective two-arm telepathology study to clinically validate SRH for telepathology at our institution. Using surgical specimens from 47 subjects, we generated a data set composed of 47 SRH images and 47 matched whole slide images (WSIs) of formalin-fixed, paraffin-embedded tissue stained with hematoxylin and eosin, with associated intraoperative clinicoradiologic information and structured diagnostic questions. We compared diagnostic concordance between WSI and SRH-rendered diagnoses. Also, we compared the 1-year median turnaround time (TAT) of intraoperative conventional neuropathology frozen sections with prospectively rendered SRH-telepathology TAT. All SRH images were of sufficient quality for diagnostic review. A review of SRH images showed high accuracy in distinguishing glial from nonglial tumors (96.5% SRH vs 98% WSIs) and predicting final diagnosis (85.9% SRH vs 93.1% WSIs). SRH-based diagnosis and WSI-permanent section diagnosis had high concordance (κ = 0.76). The median TAT for prospectively SRH-rendered diagnosis was 3.7 minutes, approximately 10-fold shorter than the median frozen section TAT (31 minutes). The SRH-imaging procedure did not affect ancillary studies. SRH generates diagnostic virtual histologic images with accuracy comparable to conventional hematoxylin and eosin-based methods in a rapid manner. Our study represents the largest and most rigorous clinical validation of SRH to date. It supports the feasibility of implementing SRH as a rapid method for intraoperative diagnosis complementary to conventional pathology laboratory methods.

Acute anoxic changes in peripheral nerve: anatomic and physiologic correlations.

INTRODUCTION: The response of the peripheral nerve to anoxia is modulated by many factors including glucose and temperature. The purposes of this article are to demonstrate the effects of these factors on the pathological changes induced by anoxia and to compare the electrophysiologic changes and pathological changes in the same nerves. METHODS: Sciatic nerves were harvested from rats and placed in a perfusion apparatus where neurophysiologic responses could be recorded continuously during a 16 h experiment. After the experiment, light microscopy and electron microscopy were performed. RESULTS: Light microscopic images showed mild changes from anoxia at normoglycemia. Hypoglycemic anoxia produced massive axonal swelling while hyperglycemic anoxia produced apparent changes in the myelin. Anoxic changes were not uniform in all axons. Electron microscopy showed only minor disruptions of the cytoskeleton with anoxia during normoglycemia. At the extremes of glucose concentration especially with hyperglycemia, there was a more severe disruption of intermediate filaments and loss of axonal structure with anoxia. Hypothermia protected axons from the effect of anoxia and produced peak axonal swelling in the 17-30°C range. CONCLUSIONS: The combination of hyperglycemia or hypoglycemia and anoxia produces extremely severe axonal disruption. Changes in axonal diameter are complex and are influenced by many factors.

Primary CNS T-cell lymphoma of the spinal cord: case report and literature review.

Primary central nervous system lymphoma (PCNSL) accounts for 1% of all lymphoma diagnoses and as many as 6% of all central nervous system (CNS) tumors. Most cases of PCNSL are of B-cell type; few are of T-cell lineage. PCNSL mainly occurs intracranially; primary spinal-cord lymphoma only occurs rarely. Moreover, intramedullary presentation without intracranial lesions is virtually unknown. Herein, we present a case of primary T-cell CNS lymphoma limited to the intramedullary spinal cord in an 82-year-old white man, along with a review of the literature on this condition and similar conditions.

Radiology-pathology conference. Acinar cell carcinoma of the pancreas.

Acinar cell carcinoma (ACC) is a rare tumor that constitutes 1% of pancreatic neoplasms. ACC is defined as a carcinoma exhibiting pancreatic enzyme production by neoplastic cells. Clinical presentation is usually related to either local spread or metastasis. In this Radiology-Pathology Conference, the clinical presentation and imaging findings of a patient with ACC of the pancreas, along with the differential diagnosis, are reviewed.

Clear cell meningioma of the cauda equina in an adult: case report and literature review.

In the pediatric population, clear cell meningiomas are more frequently intracranial than intraspinal in location. Tumors recur in up to 40% of cases within 15 postoperative months and are often managed with repeated resection with or without radiation therapy. The management strategy for adults with clear cell meningiomas involving the lumbar spinal canal (cauda equina) is less clearly defined. A 41-year-old woman presented with mild, right greater than left, lower extremity paresis. An enhanced magnetic resonance (MR) scan revealed a homogeneously enhancing intradural lesion filling the spinal canal at the L3-L4 level. Preoperative noncontrast MR studies of the brain and cervical and thoracic spine were negative. An L2-L5 laminectomy was performed for gross total excision of the intradural lesion, which was adherent to one nerve root of the cauda equina. Frozen-section diagnosis confirmed clear cell tumor. Differential diagnoses included meningioma versus renal cell carcinoma. Negative postoperative chest, abdominal, and pelvic computed tomography studies ruled out tumor of renal cell origin. Enhanced MR studies of the neuraxis proved negative. Consultations with multiple oncologists and radiation therapists recommended neither radiation nor chemotherapy following this initial surgery. She remains disease-free 1 year postoperatively. The high recurrence rate for clear cell meningiomas in children requires repeated tumor resection with or without secondary radiation therapy. Following gross total resection of lumbar tumors in adults, reserving radiation therapy for secondary recurrences provides optimal management.

Acute effects of nonlethal in utero hypoxia on fetal guinea pig heart and lack of persistent cardiac or cerebral effects in the neonate.

This study assesses the vulnerability of fetal guinea pig heart to metabolic changes during acute nonlethal in utero hypoxia. Guinea pigs (50-55 days gestation) were exposed to 7% O2 for 2 h and room air for 4 h. Fetal hearts were harvested before hypoxia, at the end of hypoxia, and 4 h after hypoxia, and analyzed for: apoptosis (TUNEL), histology, lipid peroxidation and ATP. A group of posthypoxic dams was taken to gestation. Within 48 h postpartum, the function of neonatal hearts was tested and cerebral histology examined. Fetal heart ATP was decreased by 27% at the end of hypoxia and by 32% 4 h after hypoxia. The lipid peroxides, 4-hydroxynonenal and malondialdehyde, were decreased by 37 and 46%, respectively, by 4 h after hypoxia. The apoptotic index increased from 2% in prehypoxic hearts to 8.4% by 4 h after hypoxia. Fetal heart morphology was unremarkable. Postpartum neonatal cardiac function was not affected and cerebral histology was unremarkable. These results support the conclusion that nonlethal in utero hypoxia has acute effects on the fetal heart but no persistent cardiac or cerebral effects in the postpartum neonate.

Safety and efficacy of intratracheal recombinant human Clara cell protein in a newborn piglet model of acute lung injury.

Despite the widespread use of exogenous surfactant, acute and chronic lung injury continues to be a major cause of morbidity in preterm infants. CC10 is a protein produced by Clara cells that inhibits phospholipase A2 and has anti-inflammatory and antifibrotic properties. We studied whether intratracheal (IT) recombinant human Clara cell protein (rhCC10) could safely minimize lung injury in a newborn piglet model of acute lung injury. Twenty-nine newborn piglets were given Survanta and then ventilated for 48 h receiving the following: room air (group 1); 100% O2 (group 2); or 100% O2 and 25, 5, or 1 mg/kg (groups 3, 4, and 5, respectively) of IT rhCC10 (diluted to 2 mL/kg with saline) at time 0. Laboratory studies, oxygen ratios, static pressure-volume curves, bronchoalveolar lavage (for inflammatory markers), and histologic analyses were performed over the 48-h study period. Pulmonary compliance and oxygenation were significantly improved in animals receiving 5 mg/kg IT rhCC10 compared with room air and 100% O2 controls (p < 0.004 and p < 0.05, respectively, ANOVA). Reductions in inflammatory markers were seen in animals receiving rhCC10, although changes did not reach statistical significance. No significant toxicity was noted. rhCC10 appeared safe and improved pulmonary function in this newborn piglet model of hyperoxic lung injury. We speculate that rhCC10 may represent a promising therapy for the prevention of lung injury in preterm infants.